Before DHPLC analysis, the target DNA fragments of interest are first amplified by PCR. Three DNA polymerases are suggested for use with DHPLC analysis, including Optimase® Polymerase, Maximase™ Polymerase and T-Taq™ Polymerase (Transgenomic).
The functional integrity of the column might be adversely affected if other DNA polymerases are used. It is also important to note that under no circumstances should PCR additives such as mineral oil, dimethyl sulfoxide, and formamide be used.
Otherwise, the column would be damaged. The amplified PCR products can be injected directly into the separation column if they are to be analyzed under the nondenaturing condition. Otherwise, they are further processed as described in the following sections on the basis of the purpose of the analysis.